Manuel Kunisch, Sascha Beutler, Christian Pilger, Friedemann Kiefer, Thomas Huser, Benedikt Wirth
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引用次数: 0
摘要
在生物体中,由心跳、呼吸或肌肉运动引起的自然运动导致组织变形,这是由组织结构的平移和拉伸引起的。这种影响会导致活体显微镜观察平面的位移或变形,并导致产生的图像数据的运动引起的像差。这反过来又严重限制了在活体成像实验中观察特定事件的时间。如果能够补偿组织运动,使观察平面保持稳定,则可以克服这些限制。我们已经开发了一个数学形状空间模型,可以预测类似血管的圆柱形组织幽灵的周期性运动。然后利用该模型快速计算出双光子激光扫描荧光显微镜的未来观测平面位置。焦平面连续调整到计算位置与一个压电驱动物镜支架。我们展示了主动运动补偿的船舶幻影的非谐波轴向位移,视场高达400 μ m × 400 μ m,垂直振幅超过100 μ m,以0.5 Hz的速率。
Active axial motion compensation in multiphoton-excited fluorescence microscopy.
In living organisms, the natural motion caused by heartbeat, breathing, or muscle movements leads to the deformation of tissue caused by translation and stretching of the tissue structure. This effect results in the displacement or deformation of the plane of observation for intravital microscopy and causes motion-induced aberrations of the resulting image data. This, in turn, places severe limitations on the time during which specific events can be observed in intravital imaging experiments. These limitations can be overcome if the tissue motion can be compensated such that the plane of observation remains steady. We have developed a mathematical shape space model that can predict the periodic motion of a cylindrical tissue phantom resembling blood vessels. This model is then used to rapidly calculate the future position of the plane of observation of a two-photon laser scanning fluorescence microscope. The focal plane is continuously adjusted to the calculated position with a piezo-actuated objective lens holder. We demonstrate active motion compensation for non-harmonic axial displacements of the vessel phantom with a field of view up to 400 µm × 400 µm, vertical amplitudes of more than 100 µm, and at a rate of 0.5 Hz.
期刊介绍:
Optics Express is the all-electronic, open access journal for optics providing rapid publication for peer-reviewed articles that emphasize scientific and technology innovations in all aspects of optics and photonics.